张强,罗彩云,孙达权,曾志锐,郑志菊,徐国强
ZHANG Qiang,LUO Caiyun,SUN Daquan,ZENG Zhirui,ZHENG Zhiju,XU Guoqiang

• 1:贵州医科大学基础医学院
• 2:贵阳市第二人民医院手术室
• 3:贵州医科大学第三附属医院医学研究中心

摘要(Abstract)：

目的 探究Nedd4家族交互作用蛋白2(Ndfip2)对人肝癌细胞SMMC-7721和HepG2增殖和凋亡能力的影响。方法 选择正常肝细胞LO2、肝癌细胞SMMC-7721和HepG2进行培养，采用蛋白印迹法检测3组细胞中的Ndfip2的蛋白表达；对培养的SMMC-7721和HepG2细胞转染靶向Ndfip2小干扰RNA(siRNA)构建Ndfip2基因沉默组（si-Ndifip2组），转染无义siRNA序列构建阴性对照组（NC组），采用蛋白印迹检测两组细胞的转染效率，CCK-8、平板克隆形成和流式细胞术检测两组细胞的增殖率和凋亡率，蛋白免疫印迹检测相关细胞内磷酸酶及张力蛋白同源的基因（PTEN）、磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B(Akt)、磷酸化Akt (Ser473)[p-Akt(S473)]、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、Bcl2关联X蛋白（Bax）、B淋巴细胞瘤-2(Bcl2)的蛋白表达。结果 与正常肝细胞LO2组比较，肝癌细胞SMMC-7721组和HepG2组中Ndfip2表达升高（P<0.05）；与NC组比较，si-Ndfip2组SMMC-7721和HepG2细胞Ndfip2蛋白表达明显下调（P<0.01）；与NC组相比，siNdifip2组肝癌细胞的增殖速率明显降低（P<0.01），凋亡率增加（P<0.05）;si-Ndifip2组肝癌细胞中PI3K、pAkt(S473)和Bcl2的蛋白表达水平均下降（P<0.05）；而PTEN、E-cadherin、Caspase-3和Bax蛋白的表达则增加（P<0.05）。结论 沉默Ndfip2抑制肝癌细胞的增殖能力，促进肝癌细胞凋亡，其可能机制与PTEN-PI3K/Akt信号通路的失活有关。
Objective To explore the effect of the expression of Nedd4 family interacting protein 2(Ndfip2) on the proliferation and apoptosis of human hepatocellular carcinoma cell( HCC) lines SMMC-7721 and HepG2.Methods The normal liver cell line LO2, HCC lines SMMC-7721,and HepG2 were selected for cell culture. The protein expression level of Ndfip2 was detected by Western blot. SMMC-7721 and HepG2 cell lines were transfected with siRNA against Ndifip2( si-Ndifip2group) or nonsense siRNA as a negative control(NC group), respectively. Silencing efficiency was verified using Western blot. CCK-8 assay, clonogenic formation assay, and flow cytometry were used to detect cell viability, cell proliferation and apoptosis rate. Western blot was used to detect the protein expression levels of phosphatase and tensin homolog(PTEN), phosphatidylinositol 3-kinase(PI3K),and protein kinase B(Akt), phosphor-Ser473-Akt [p-Akt(S473)], Caspase-3, Bcl2-associated X protein( Bax), B lymphoma-2( Bcl2).Results When compared to normal liver cell line LO2,Ndfip2 expression was significantly increased in cell lines SMMC-7721 and HepG2(P< 0. 05).siRNA against Ndfip2 significantly downregulated Ndfip2 protein expression in si-Ndifip2 group relative to NC group(P< 0. 01). When compared to the NC group, cell proliferation rates of si-Ndifip2 groups were significantly reduced(P< 0. 01), while their apoptosis rates were significantly increased(P<0. 05). When compared to the NC group, the protein expression levels of PI3K,p-Akt(S473), and Bcl2 were decreased significantly(P< 0. 05), while the expressions levels of PTEN, E-cadherin,Caspase-3, and Bax were significantly increased(P< 0. 05).Conclusion Ndfip2 silencing-triggered cell proliferation inhibition and apoptosis induction of HCC cells may be related to the inactivation of the PTEN-PI3K/Akt signaling pathway.

关键词(KeyWords)： 肝肿瘤细胞;Nedd4家族交互作用蛋白2;增殖;凋亡;基因;蛋白激酶B
hepatocellular carcinoma(HCC) cells;Nedd4 family interacting protein 2(Ndfip2);cell proliferation;apoptosis;gene;protein kinase B(Akt)

Abstract:

Keywords:

基金项目(Foundation): 贵州省中医药、民族医药科学技术研究课题（黔中医药发[2015]8,QZYY-2015-012）;; 贵州省卫生计生委科学技术基金项目（gzwjkj2014-1-029）

作者(Author): 张强,罗彩云,孙达权,曾志锐,郑志菊,徐国强
ZHANG Qiang,LUO Caiyun,SUN Daquan,ZENG Zhirui,ZHENG Zhiju,XU Guoqiang

DOI: 10.19367/j.cnki.2096-8388.2022.04.005

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